6-FOLD ROTATIONAL SYMMETRY OF CLPQ, THE ESCHERICHIA-COLI HOMOLOG OF THE 20S-PROTEASOME, AND ITS ATP-DEPENDENT ACTIVATOR, CLPY

ClpQ (HslV) is a homolog of the beta-subunits of the 20S proteasome. I n E. coli, it is expressed from an operon that also encodes ClpY (HsIU ), an ATPase homologous to the protease chaperone, ClpX. ClpQ (subunit M(r) 19 000) and ClpY (subunit M(r) 49 000) were purified separately as oligomeric proteins with molecular weights of similar to 220 000 an d similar to 350 000, respectively, estimated by gel filtration. Mixtu res of ClpY and ClpQ displayed ATP-dependent proteolytic activity agai nst casein, and a complex of the two proteins was isolated by gel filt ration in the presence of ATP. Image processing of negatively stained electron micrographs revealed strong six-fold rotational symmetry for both ClpY and ClpQ, suggesting that the subunits of both proteins are arranged in hexagonal rings. The molecular weight of ClpQ combined wit h its symmetry is consistent with a double hexameric ring, whereas the data on ClpY suggest only one such ring. The symmetry mismatch previo usly observed between hexameric ClpA and heptameric ClpP in the relate d ClpAP protease is apparently not reproduced in the symmetry-matched ClpYQ system.